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Morpho-Physiological Processes in Embryonic Development of Odessa Barb Puntius Padamya Kullander & Britz, 2008 (Teleostei: C

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Morpho-Physiological Processes in Embryonic Development of Odessa Barb Puntius Padamya Kullander & Britz, 2008 (Teleostei: C MORPHO-PHYSIOLOGICAL PROCESSES IN EMBRYONIC DEVELOPMENT OF ODESSA BARB PUNTIUS PADAMYA KULLANDER & BRITZ, 2008 (TELEOSTEI: CYPRINIDAE) IN THE OPTIMAL AND REDUCED PH Agata Korzelecka-Orkisz Agata Gajewska Adam Tański Krzysztof Formicki Department of Hydrobiology, Ichthyology and Biotechnology of Reproduction, The Westpomeranian University of Technology in Szczecin, Poland [email protected] Abstract This study describes the structures of ruby barb (Puntius padamy), morpho- physiological changes during embryogenesis from activation to hatching under optimal conditions (25°C; pH 7.0), the effect of acidic pH (5.8) on the early developmental stages of the species, the relationship between pH water and fry survival. Water pH quite significantly influenced the rate of embryogenesis – the differences in the rate of development had already emerged at the stage of eight blastomeres. The total duration of embryogenesis from fertilization to the moment of mass hatching of larvae under pH 7.0 was 650H (27 hours) and under pH 5.8 was 960H (40 hours). The percentage of hatching inside pH 5.8 water was 10%, but 60% inside pH 7.0 water. Water pH quite significantly affected somatic motility of embryos. Keywords pH, spawning, ruby barb, Puntius padamya INTRODUCTION The odessa barb Puntius padamya, also known as "ruby barb" is one of the most interesting fish of the carp family, popular in the aquarium cultures. Its origin is not fully understood (Dazkewitsch 1973a, 1973b, Stallknecht 1973, Mills 2002, Arunkumar and Tombi Singh 2003, Axelrod et al.. 2008). Most probably it was presented for the first time on Odessa fish bazaar about 1971 - 1972. Its name - "Odessa barb" originates precisely from the city of Odessa (Hochstrasser 1980, Nieuwenhuizen 1984, Sterba 1988, Menon i in. 2000, Vishwanath and Laisram 2004; 12 Kullander and Fang 2005, Rüber i in. 2007). Kullander and Britz in 2008 described it as a new species (Kullander 2008, Kullander and Britz 2008,). Odessa barb is a quiet gregarious fish. Adults grow to a length of up to 8 cm (Frey 1990). The body is of pale-yellow silver color, two dark spots run along its sides: one just behind the gillnet, the second above the anal fin (Mills 2002). All fins of the odessa barbs are have an orange-red hue (Krzykawski et al., 2001). The fish tolerates a wide temperature range, 14-25C, however, only at 20C are the specimens fully stained (Frey 1990, Dreyer and Keppler 1996, Kahl et al.in. 1997). Sexual dimorphism in odessa barb is clearly marked. Mature males have a broad red band along the side of the body, they are more slender and have "dotted" fins (Frey 1990). Coloration of the males becomes more intense during spawning. Females during the breeding readiness period have the belly more prominent (Mills 2002). Fish rub themselves most often among tiny illuminated cirriform vegetation (preferably in the morning sun). After superb chase, they freely release the eggs and this can take several hours because the barbs spawn in batches. Individual fertility is about 150 eggs (Frey 1990, Talwar and Jhingran 1991, Pertovicky 2003). The yellowish spawn is equipped with a sticky cover, with the help of whose it attaches itself to the vegetation. Development takes about 24 hours at 25C. The larvae have glands that secrete sticky substance allowing it to attach itself to items such as tank wall, and after about 5 days, they begin to actively swim (Talwar and Jhingran 1991, Petrovicky 2003). Their reproductive strategy is very similar to that adopted by the closely related species of cherry barb Puntius titteys. This species also lays eggs on aquatic vegetation and this is facilitated by the stickiness of the casing. Development of cherry barbs takes 19-36 hours, depending on temperature. The average volume of the cherry barbs egg is 0.97 ± 0.09, and yolk sphere inside them measure, on average, 0.40 ± 0.06. Large perivitellar space constitutes, which creates favorable conditions for the embryo to breath and allows for free movement, makes up to 60% of the egg volume. After about 1.5 h of development, thick-cell morula appears, closing of blastopor takes place after 4.5 hours of development. Like in most carp, the yolk sac in cherry barbs is 13 divided into two branches: the bigger front branch known as proximal and the smaller rear known as caudal. The first somatic contractions are observable after just some 10 hours from activation, after 14.5 h the heart starts operating and attain a rate of 72 beats per minute just before hatching (Korzelecka-Orkisz et al 2009b). The pH of water has a highly significant effect on living organisms. Environmental acidification resulting inter alia from anthropogenic pollution is noticeable in almost every corner of the world. Study on the impact of low-pH water on the juvenile stages of herring (Clupea harengus L.) have showed that although no linear correlation between the level of acidification and the total length, weight, yolk sac size and size of otoliths of newly hatched larvae was demonstrated, however, concentration of RNA at hatching was reduced, which consequently resulted in lower concentrations of protein and thus may lead in future to reduced weight of developing fish (Franke and Clemmesen 2011). Also, in fish of the Lebistes reticulatus species subjected to a 24-hour exposure to water of pH 4.11-4.39, the result was modification of fish metabolism, which was manifested by an increase in lipid content of faces, by 4-36% per mg of dry weight, as compared with the energy content of food. This phenomenon occurred only in juveniles (Urban-Jezierska 2002). Since it is difficult to predict the reaction of fish and other aquatic organisms to changing water pH, an attempt was made to draw a possible scenario for the next 50- 100 years by subjecting the orange clownfish, Amphiprion percula to the actions of these factors. Acidification did not affect the duration of the embryogenesis, eggs survival rate and brood size of this species, however, it did have an impact on the size and weight of larvae – larvae were 15 to 18 per cent longer and 47 to 52 per cent heavier in acidified water compared with those in control setting (Munday at al. 2009). In Germany, many pits left over from brown coal mines were flooded, and because residues from mining affected the water pH, therefore, the range of tolerance at juveniles stages of the Tinca tinca was examined. All embryos in water of pH 3.50- 4.75 died before hatching or shortly after hatching. In water of pH 5.50, only 3 - 4% survived, which was taken as the threshold limit which can be tolerated by Tinca tinca embryo (Duis 2001). 14 Adult fish are better able to adapt to unfavorable environmental conditions caused by low water pH (Brown and Sadler 1989), although sensitivity of species to changing pH varies. Lowering water pH to 6.0, in the absence of other adverse factors, generally do not cause any adverse effects on fish organism. Temporal further reduction of pH by 1 degree does lead to noticeable, permanent damage, although maintaining lower pH for a long period would damage delicate structures of the gills, which reduces the possibility of oxygen uptake by this organ. At pH below 5.0, they are less mobile and tend to swim to the surface. At this water pH, symptoms of acid effects can already be observed – skin are covered with large amounts of mucus while gills are darker. Acidification increases the susceptibility of fish to other diseases and reduces their rate of development (Kocyłowski and Miączyński 1960). Although odessa barb is a popular aquarium fish and is often used in laboratory experiments (Prakash and Kapoor 1976, Taki 1978, Satyanarayan and Ramakant 2004, Sahoo et al.. 2007, Sevilla 2007), there is no literature on both embryogenesis and impact in which adverse environmental conditions, including reduced pH, has on this fish. This has aroused our interest, hence we decided to follow the process of embryogenesis under optimal conditions and pH, compared to conditions under slightly reduced pH, and on this basis try to explain biological sense of observed differences, and show relationship existing between specific morpho-mechano- physiological processes and the chemical parameters (water acidic) of the environment in which natural reproduction takes place. During the experiments, changes taking place in the egg of developing embryo were observed and recorded, the heart rate and embryonic motility were monitored, in addition, the state of newly hatched specimens was analyzed. MATERIALS AND METHODS Studies on embryonic development of the odessa barb Puntius padamya were carried out from April 2007 to May 2009, in an aquarium room belonging to the Department of Hydrobiology, Ichthyology and Biotechnology of Reproduction ZUT in Szczecin. Preparation of material - material for testing were obtained from specimens that had been breed from juvenile stage to sexual maturity. After 7 months of pre- 15 rearing, the best specimens were selected (10 males and 7 females). Prior to breeding, spawners were conditioned for 7 days. Water of adequate physical and chemical parameters was prepared 48 hours before releasing spawners into a 25-liter glass spawning aquarium, with a 1:1 ratio of distilled water to tap water, aerated for 72 hours (water temperature - 25 ±05C, water pH - 7 ± 0.5, total hardness - 8n). After spawning, the adults were transferred to breeding aquarium, while the eggs were transferred to a 0.5-liter mini aquariums with water parameters as shown in Tab.1 Table 1. Experimental treatments Temperature Water Hardness [C] pH of water [n] Variant no 1 25±0,5 7,0±0,5 8 (control) Variant no 2 25±0,5 5,8± 0,2 8 Embryonic and larval development The water in the tanks containing the spawn was constantly aerated, and the lower pH was maintained using Tropical-pH minus ( Tropical Company), a medium used in aquarium.
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